Research Article

Gender Difference on the Effect of Omega-3 Polyunsaturated Fatty Acids on Acetaminophen-Induced Acute Liver Failure

Figure 3

n-3 PUFA-modulated hepatic oxidative response and autophagy activation against APAP toxicity is distinct between male and female mice. (a–d) Male or female WT and fat-1 mice were injected with 400 mg/kg APAP, and livers were collected at 6 hours post-APAP injection (). (a) Flow cytometry analysis of the intercellular ROS level by the fluorescent probe DCFH-DA in hepatocytes was carried out. (b) The mitochondrial membrane potential in hepatocytes was measured by the JC-1 dye staining for flow cytometry analysis. (c) Phosphorylation of JNK was evaluated by immunoblotting analysis at the indicated time after APAP challenge. (d) The protein levels of LC3 and p62 were examined by immunoblotting analysis. (e–g) HepaRG cells were pretreated with 50 μM DHA with or without100 nM E2 prior to stimulation with 20 mM APAP. (e) The ROS level in the cells was analyzed by flow cytometry labeling with fluorescent probe DCFH-DA at 6 hours following APAP administration. (f) Phosphorylation of JNK expression in the APAP-treated cells was evaluated by immunoblotting analysis. (g) The hepatic levels of LC3 and p62 were determined by immunoblotting analysis. , . The data represent three independent experiments with similar results.